Primer for detecting trichothecene group B-class toxin of fusaria through PCR and application of primer

A technique for detecting trichothecenes and primers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problems of cumbersome sample purification, high technical content requirements, and low cost. , to achieve the effects of easy large-scale promotion and application, accurate detection results, and simple operation

Active Publication Date: 2015-04-29
南京微测生物科技有限公司
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Problems solved by technology

[0004] At present, the detection methods of mycotoxin content usually include biological detection methods, physical and chemical detection methods, immunochemical detection methods, etc. Due to the lack of uniform standardized standards and inaccurate detection and analysis of biological detection methods, they have not been used in the actual detection of fusarium toxins. It can be widely used; the chemical determination method is currently the main method used for the determination of Fusarium toxin content, which is characterized by accuracy and good repeatability, but the sample purification required by this method is cumbersome and requires high technical content, which cannot meet the requirements of rapid detection. Requirements; immunoassay, simple operation, low cost, but because the detected toxin often contains some other derivatives, so the result of the determination is often inaccurate, prone to false positives; these three detection methods are not suitable for rapid, Sensitive and Accurate Detection of Class B Trichothecenes Contamination
[0005] In recent years, with the publication of a variety of Fusarium genomes and the rapid development of molecular biology research, researchers at home and abroad have made important achievements in the synthesis of trichothecene toxins and the regulation of toxin synthesis gene expression, confirming that The important role of some genes in the pathway of toxin metabolism has been discovered. Based on these research results and the sequence information of related genes, establishing a simple, rapid and inexpensive toxin detection method has become a hot spot for researchers from different countries. Kim et al. used Tri5, Tri7 and Tri13 gene sequences, and detected the production of Fusarium graminearum DON and NIV toxins from multiple hosts (Kim et al.Polymorphism of trichothecene biosynthesis...

Method used

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  • Primer for detecting trichothecene group B-class toxin of fusaria through PCR and application of primer
  • Primer for detecting trichothecene group B-class toxin of fusaria through PCR and application of primer
  • Primer for detecting trichothecene group B-class toxin of fusaria through PCR and application of primer

Examples

Experimental program
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Effect test

Embodiment 2

[0031] Example 2 Detection of Fusarium Trichothecenes B Toxoids in Wheat and Corn Grains

[0032] Several samples of wheat and corn with natural disease in the field were taken, ground and pulverized, and template DNA was extracted. The extraction method was the same as in Example 1. The types and contents of toxins contained in the samples were shown in Table 2.

[0033]

[0034] PCR reaction is identical with embodiment 1 with electrophoresis detection method, and detection result is as follows figure 2 As shown, the M lane is the standard value, and its lane numbers correspond to the sample numbers one by one. Among them, the wheat samples in lanes 1 and 2 amplified two specific bands of 443bp and 376bp, and the wheat samples in lanes 3 and 4 In the sample, three specific bands of 443bp, 376bp and 246bp were amplified, and in the corn samples of lanes 5 and 6, two specific bands of 443bp and 376bp were amplified.

[0035] The toxin-producing types of samples 1-6 were v...

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Abstract

The invention discloses a set of primer for detecting trichothecene group B-class toxin of fusaria through PCR (polymerase chain reaction). The primer comprises a share sense primer F shown in SEQID NO.1, 3-acetyl deoxygenation nivalenol reverse primer R1 shown in SEQID NO.2, 15-acetyl deoxygenation nivalenol reverse primer R2 shown in SEQID NO.3 and nivalenol reverse primer R3 shown in SEQID NO.4; the set of the primers can be applied to the PCR for directly detecting 3-acetyl deoxygenation nivalenol, 15-acetyl deoxygenation nivalenol and nivalenol. The primer is low in cost, simple in operation, short in consumption time, accurate in testing result and easy for large-scale popularization and application.

Description

technical field [0001] The invention belongs to the field of food safety detection, in particular to PCR detection primers for fusarium trichothecene family B toxoids and applications thereof. Background technique [0002] Fusarium head blight (or scab) caused by Fusarium is an important epidemic disease worldwide, which can harm many cereal crops including barley, wheat, corn, rice and so on. With global climate warming and abnormal regional climate and changes in farming systems, the frequency of disease occurrence has increased and the area of ​​occurrence has continued to expand. Fusarium scab not only leads to serious loss of grain yield, but also Fusarium can produce a variety of mycotoxins, reduce the quality of food and feed, seriously threaten the health of humans and animals, and bring major hidden dangers to food safety. [0003] Trichothecenes are the most important mycotoxins produced by Fusarium, which are mainly divided into two types: A and B. It can inhibi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/02C12Q1/6876
Inventor 仇剑波董飞徐剑宏史建荣
Owner 南京微测生物科技有限公司
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